Recent Advances and the Potential for Clinical Use of Autofluorescence Detection of Extra-Ophthalmic Tissues

The autofluorescence (AF) characteristics of endogenous fluorophores allow the label-free assessment and visualization of cells and tissues of the human body. While AF imaging (AFI) is well-established in ophthalmology, its clinical applications are steadily expanding to other disciplines. This review summarizes clinical advances of AF techniques published during the past decade. A systematic search of the MEDLINE database and Cochrane Library databases was performed to identify clinical AF studies in extra-ophthalmic tissues. In total, 1097 articles were identified, of which 113 from internal medicine, surgery, oral medicine, and dermatology were reviewed. While comparable technological standards exist in diabetology and cardiology, in all other disciplines, comparability between studies is limited due to the number of differing AF techniques and non-standardized imaging and data analysis. Clear evidence was found for skin AF as a surrogate for blood glucose homeostasis or cardiovascular risk grading. In thyroid surgery, foremost, less experienced surgeons may benefit from the AF-guided intraoperative separation of parathyroid from thyroid tissue. There is a growing interest in AF techniques in clinical disciplines, and promising advances have been made during the past decade. However, further research and development are mandatory to overcome the existing limitations and to maximize the clinical benefits

Digital Image Analysis of Vitiligo for Monitoring of Vitiligo Treatment

Vitiligo is an acquired pigmentary skin disorder characterized by depigmented macules that result from damage to and destruction of epidermal melanocytes. Visually, the vitiligous areas are paler in contrast to normal skin or completely white due to the lack of pigment melanin. The course of vitiligo is unpredictable where the vitiligous skin lesions may remain stable for years before worsening. Vitiligo treatments have two objectives, to arrest disease progression and to re-pigment the vitiligous skin lesions. To monitor the efficacy of the treatment, dermatologists observe the disease directly, or indirectly using digital photos. Currently there is no objective method to determine the efficacy of the vitiligo treatment. Physician's Global Assessment (PGA) scale is the current scoring system used by dermatologists to evaluate the treatment. The scale is based on the degree of repigmentation within lesions over time. This quantitative tool however may not be help to detect slight changes due to treatment as it would still be largely dependent on the human eye and judgment to produce the scorings. In addition, PGA score is also subjective, as it varies with dermatologists. The progression of vitiligo treatment can be very slow and can take more than 6 months. It is observed that dermatologists find it visually hard to determine the areas of skin repigmentation due to this slow progress and as a result the observations are made after a longer time frame. The objective of this research is to develop a tool that enables dermatologists to determine and quantify areas of repigmentation objectively over a shorter time frame during treatment. The approaches towards achieving this objective are based on digital image processing techniques. Skin color is due to the combination of skin histological parameters, namely pigment melanin and haemoglobin. However in digital imaging, color is produced by combining three different spectral bands, namely red, green, and blue (RGB). It is believed that the spatial distribution of melanin and haemoglobin in skin image could be separated. It is found that skin color distribution lies on a two-dimensional melanin-haemoglobin color subspace. In order to determine repigmentation (due to pigment melanin) it is necessary to perform a conversion from RGB skin image to this two-dimensional color subspace. Using principal component analysis (PCA) as a dimensional reduction tool, the two-dimensional subspace can be represented by its first and second principal components. Independent component analysis is employed to convert the twodimensional subspace into a skin image that represents skin areas due to melanin and haemoglobin only. In the skin image that represents skin areas due to melanin, vitiligous skin lesions are identified as skin areas that lack melanin. Segmentation is performed to separate the healthy skin and the vitiligous lesions. The difference in the vitiligous surface areas between skin images before and after treatment will be expressed as a percentage of repigmentation in each vitiligo lesion. This percentage will represent the repigmentation progression of a particular body region. Results of preliminary and pre-clinical trial study show that our vitiligo monitoring system has been able to determine repigmentation progression objectively and thus treatment efficacy on a shorter time cycle. An intensive clinical trial is currently undertaken in Hospital Kuala Lumpur using our developed system. VI

Rosacea as a Disease of Cathelicidins and Skin Innate Immunity

Rosacea is a common and chronic inflammatory skin disease most frequently seen in groups of genetically related individuals. Although the symptoms of rosacea are heterogeneous, they are all related by the presence of characteristic facial or ocular inflammation involving both the vascular and tissue stroma. Until recently, the pathophysiology of this disease was limited to descriptions of a wide variety of factors that exacerbate or improve disease. Recent molecular studies show a common link between the triggers of rosacea and the cellular response, and these observations suggest that an altered innate immune response is involved in disease pathogenesis. Understanding rosacea as a disorder of innate immunity explains the benefits of current treatments and suggests new therapeutic strategies for alleviating this disease

Physical anthropology in Japan: The Ainu and the search for the origins of the Japanese

In this paper I examine the quest by physical anthropologists in Japan for the origins of the Japanese. A major focus of this research has been the Ainu people of the northern island of Hokkaidō, who have recently been declared an indigenous people of Japan. The relationship between mainstream Japanese and the very much living community of the Ainu has been the subject of over 100 years of research. Integral to research has been the collection of Ainu skulls, skeletons, and artefacts that have provided a critical if controversial resource for physical anthropologists. This has all been against the backdrop of changing political ideologies about the so-called purity of the Japanese. In the post–World War II period, with the loss of empire, the idea of Japan as a homogeneous nation took hold, and it was only in the last two decades that this notion has been discredited

Studies of different clinical manifestations of Sarcoidosis and the role of genetic factors

Sarcoidosis is a systemic disease of unknown etiology characterized by the formation of non-necrotizing granulomas in the affected organs. Engagement of the lungs and/or thoracic lymph nodes (LN) are found in more than 90 % of all cases, but almost any organ such as the eyes, skin, heart and nervous system can be involved. Genetic factors influence the risk for disease as well as the clinical picture seen in sarcoidosis and especially the genes localized to the human leukocyte antigen (HLA) region on chromosome six are believed to be of importance. For example, the HLA-DRB1*0301 allele is found to be strongly associated with Löfgren’s syndrome (LS). Characteristic for LS is an acute onset usually with fever, bilateral ankle arthritis and/or erythema nodosum and bilateral hilar lymphadenopathy with in some cases parenchymal infiltrates. The HLA-DRB1*0301 allele is also associated with an accumulation of T cells expressing the T cell receptor variable gene segment AV2S3 in bronchoalveolar lavage fluid (BALF) of sarcoidosis patients. The aim of this thesis has been to identify risk factors for different clinical manifestations in sarcoidosis as well as markers of importance for the inflammatory cell response seen in sarcoidosis. The results show that HLA-DRB1*04 positive sarcoidosis patients had an increased risk for the three organ engagements associated with Heerfordt´s syndrome. Heerfordt´s syndrome is a phenotype of sarcoidosis that in its complete form consists of uveitis, parotid and/or salivary gland enlargement and cranial nerve palsy. In comparison to BALF where a high CD4/CD8-ratio is strongly associated with sarcoidosis, the CD4/CD8-ratio in the affected LNs of sarcoidosis patients had no diagnostic value. Further, in HLA-DRB1*03 positive patients the associated accumulation of AV2S3+ T cells was strictly compartmentalized in BALF. This finding indicates an airborne antigen as the triggering factor in sarcoidosis. The risk for cardiac sarcoidosis (CS) was significantly higher in patients with an abnormal electrocardiography (ECG) compared to those with a normal ECG. The risk for CS was highest in patients who had a pathologic ECG in combination with cardiac related symptoms. Further, non-LS was associated with an increased risk for CS. In LS patients was the absence of HLA-DRB1*03 a risk factor for extra-pulmonary manifestations (erythema nodosum and ankle arthritis excluded). Another risk marker in all patients was HLA-DRB1*04/*15 where half of the patients had extra-pulmonary manifestations. In conclusion, the HLA-DRB1*04 allele is associated with an increased risk for involvement of the eyes, parotid and/or salivary glands and cranial nerves in sarcoidosis patients. Moreover, an increased CD4/CD8-ratio in enlarged LNs is not diagnostic for sarcoidosis in comparison to BALF where a high ratio is strongly associated with sarcoidosis. Further, a pathologic ECG is a risk marker for CS in sarcoidosis patients. Finally, not only the single HLA-DRB1 alleles are of importance for the risk of extra-pulmonary manifestations in sarcoidosis, but also the allele combinations and where especially the combination HLA-DRB1*04/*15 calls for an increased awareness and a more intensive follow-up

Toxicology of Asparagus laricinus in rats

Published ThesisTraditional medicine has been a fertile source of revealing lead novel molecules which are then subjected to investigations using the techniques of the modern drug discovery. There are a number of conventional drugs that originate from plants, such as aspirin (from willow bark), digoxin (from foxglove), quinine (from cinchona bark), and morphine (from the opium poppy). The aim of the present work was to evaluate the possible toxic effects of the dried roots of Asparagus laricinus extracts using Sprague Dawley rats as animal models. In this study we investigated the use of Asparagus laricinus roots extracts for novel anticancer drug development, specifically looking at the safety and toxicology. Previous in vitro studies on Asparagus laricinus extracts have demonstrated anticancer activity against three human cell lines, namely, breast MCF7, renal TK10 and melanoma UACC62. These necessitated further studies on Asparagus laricinus extracts, such as toxicity, adverse effects investigations as well as in vivo biological studies using animal models. The objectives of the study was to evaluate variations in serum biochemical parameters, investigate possible deviations in haematological parameters, and also to assess histopathological variations on the liver, kidneys and spleen tissues of animals exposed to aqueous and ethanolic extracts of Asparagus laricinus roots. The study was conducted at the Animal Research Unit at the University of the Free State, Bloemfontein, South Africa.Written approval for the final version of the protocol was obtained from the Interfaculty Animal Ethics Committee of the Faculty of Health Sciences at the University of the Free State. The dried plant roots were pulverized, 10g soaked in ethanol or distilled water and agitated for 72 hours at 120 rpms. The supernatant was filtered and the ethanol removed completely under vacuum. The aqueous sample was lyophilized to obtain dried powdered material. The powdered plant material was dissolved in distilled water to prepare 2%, 10% and 20% concentration. The material was also dissolved in ethanol and different concentrations were obtained by varying the volumes of the solution administered. The study population consisted of a total of 54 Sprague Dawley rats (weighing between 180g and 250g), both male and female, obtained from the above research unit. The animals were divided into two groups of 24 and 30 rats for aqueous and ethanolic extracts, respectively. The aqueous group was further divided into four subgroups of 6 rats which were exposed to 2%, 10% and 20% extracts and the control group (unexposed). The ethanolic group was divided into five subgroups of 6 rats which were exposed to increasing doses of 50, 100, 200 and 400mg/kg/day extracts and the last group served as controls (unexposed). The aqueous extracts were administered to the three subgroups for eight weeks ad libitum while the control group was exposed to tap water. Ethanol extracts were administered daily over a period of two weeks through gavage and the control group was administered water through gavage as well. Blood samples were collected, animals were sacrificed and organs/tissues excised for histological assessment. Biochemical and haematological tests were selected as indicators of the damage to the tissue of organs, including the liver, kidney and spleen. A significant difference (p<0.05) was observed for platelets with the ethanol extract at a dose of 50g/kg/day. There were no statistical differences between the treatment groups and controls with regard to the rest of haematological variables and selected biochemical tests. Comparison of the controls (n=6) and treatment groups (n=6) revealed an average median change in weight of slightly above 50g over the entire eight-week period of experimentation with aqueous extracts. A significant difference (p<0.05) was observed for both haemoglobin and blood urea nitrogen results with the 20% water extract. There were no statistical differences between the treatment and control groups with regard to the rest of haematological variables and selected biochemical tests. Histological evaluation could not reveal any pathological changes in both the aqueous and ethanolic extracts across all levels of dosage. The main purpose of the study was to establish whether Asparagus laricinus has any toxic or adverse effects on the tissue and organs of animal models. This was done by evaluating hepatotoxicity, nephrotoxicity, spleen and vascular damage to the animals. Dose-response assessment of the effect of the extract was done by analysis of the blood and tissue samples collected at the end of the research. Biochemical and haematological results could not show any patterns in abnormalities although we observed statistically significant results on few parameters. Histologically, no pathological changes were observed. In conclusion, we summarize that the toxicological evaluation of Asparagus laricinus extracts may be considered relatively free of toxicity when given orally, as it did not cause death, damage or inflammation to the tissues, nor produced any remarkable biochemical and haematological adverse effects in both the male and female Sprague Dawley rats

Role of galectin-3 in bone cell differentiation, bone pathophysiology and vascular osteogenesis

Galectin-3 is expressed in various tissues, including the bone, where it is considered a marker of chondrogenic and osteogenic cell lineages. Galectin-3 protein was found to be increased in the differentiated chondrocytes of the metaphyseal plate cartilage, where it favors chondrocyte survival and cartilage matrix mineralization. It was also shown to be highly expressed in differentiating osteoblasts and osteoclasts, in concomitance with expression of osteogenic markers and Runt-related transcription factor 2 and with the appearance of a mature phenotype. Galectin-3 is expressed also by osteocytes, though its function in these cells has not been fully elucidated. The effects of galectin-3 on bone cells were also investigated in galectin-3 null mice, further supporting its role in all stages of bone biology, from development to remodeling. Galectin-3 was also shown to act as a receptor for advanced glycation endproducts, which have been implicated in age-dependent and diabetes-associated bone fragility. Moreover, its regulatory role in inflammatory bone and joint disorders entitles galectin-3 as a possible therapeutic target. Finally, galectin-3 capacity to commit mesenchymal stem cells to the osteoblastic lineage and to favor transdifferentiation of vascular smooth muscle cells into an osteoblast-like phenotype open a new area of interest in bone and vascular pathologies